Tumor is an abnormal lesion formed by the body's clonal dysplasia under the action of various carcinogenic factors, local tissue cells lose their normal regulation at the gene level. The academic world divides it into two categories: benign and malignant. As one of the important causes of death from cancer in the world, if the tumor is found to be ineffective, it may lead to weight loss, weakness, anemia, loss of appetite, fever and impaired organ function. The consequences are extremely serious.
Real-Time PCR (Real-Time PCR) technology is a nucleic acid quantification technology developed on the basis of PCR technology. It was introduced by Applied Biosystems in the United States in 1996 by adding a fluorescent group to the PCR reaction system and then using fluorescence. The accumulation of signals is used to monitor the entire PCR process in real time. Finally, the unknown DNA template is quantified by the standard curve, which realizes a qualitative leap to quantitative leap, providing a new and rapid and accurate analysis of various substances in the sample. The technical tools are now widely used in the detection of laboratory and clinical tumor markers.
Compared with traditional PCR technology, Real-Time PCR uses fluorescent probes to label specific nucleic acids to improve accuracy; sensitivity is increased, so fewer sample sizes are used, which reduces the risk of sample exhaustion, in an hour or so. The results can be simplified, the traditional PCR method is simplified; the closed-loop analysis and the real-time dynamic detection and automatic quantification of PCR amplification products by computer analysis software, the quantitative dynamic range is up to five orders of magnitude, and the results are reproducible. This also increases the sensitivity and precision of the test.
Real-Time PCR for tumor detection and treatment:
- Early diagnosis, typing
Ovarian cancer is the most malignant tumor with gynecological mortality. It has the characteristics of occult occult and rapid progress. Only 25% of ovarian cancer can be found in stage I lesions. Most patients have spread or metastasized at the time of diagnosis. Early diagnosis of ovarian cancer plays an important role in patients with ovarian cancer. Human epididymis protein 4 (HE4) is a new gene discovered in recent years. A large number of studies have shown that HE4 is an excellent biomarker for identifying benign and malignant ovarian lesions. The researchers used Real-Time PCR to detect advanced ovarian cancer. After surgery and treatment, HE4 increased in 4 to 5 months before recurrence, suggesting that HE4 can be used to detect recurrence of ovarian cancer.
Many researchers believe that breast cancer is a local manifestation of a systemic disease, and its therapeutic effect depends on whether it is distantly metastatic at the time of diagnosis. Metastasis and recurrence are the main causes of breast cancer death. If micrometastasis can be detected in a subclinical state, it has an important guiding role in clinical diagnosis, treatment and prognosis. The prediction and prevention of early breast cancer recurrence and metastasis are the frontier topics in breast cancer research, especially the detection of peripheral blood circulation tumor cells (CTT) and axillary lymph node micrometastasis has gradually become the focus of breast cancer research. Among the common landmark proteins are mammary epithelial small mucin (SBEM), cytokeratin 19 (CK19) and mammary gland cell (hMAM).
Studies have shown that miR-122 is closely related to hepatocellular carcinoma typing, and inhibition of miR-122 expression is a feature of liver cancer, which is characterized by enhanced invasion, tumor recurrence and reduced survival time of patients; simultaneous use of Real-Time PCR The detection of miR-122 expression by immunohistochemistry and other methods plays an important role in the typing of hepatocellular carcinoma.
- Judging prognosis
Fluorescence quantitative Real-Time PCR technology is used to evaluate the sensitivity of chemotherapy by detecting the dynamic changes of tumor target mRNA in peripheral blood during chemotherapy. The clinical recurrence and mortality of patients with positive CK19 mRNA expression after chemotherapy were significantly increased (P<0.001), while the tumor-free survival and overall survival rate were significantly lower (P<0.001). Statistical analysis showed that fluorescence quantitative Real-Time PCR detection of CK19 mRNA expression in peripheral blood before and after chemotherapy can be used as an independent indicator to determine the prognosis and overall survival rate of breast cancer patients.
The study found that for miR-429, high miR-429 expression is associated with poor prognosis in liver cancer. In recent years, there has been increasing evidence that the expression of miR-429 is associated with a variety of tumors, such as colon cancer, gastric cancer, and ovarian cancer.
- Guided treatment
The traditional clinical evaluation of chemosensitivity is mainly based on the size of the tumor and needs to be performed 3 to 4 months after the start of treatment. By detecting the dynamic changes of target mRNA in peripheral blood during tumor chemotherapy, the efficacy can be predicted as early as 3 to 4 weeks after the initial treatment, which is of great significance in determining the individualized treatment of patients, especially for patients with non-solid tumors and adjuvant chemotherapy. There are no measurable solid lesions, and this method of assessing chemosensitivity is particularly important.
In summary, Real-Time PCR is a gene quantification technology widely used in clinical practice. It can not only accurately quantify the expression of tumor-related genes, but also accurately quantify tumor markers in peripheral blood. It can be used for early diagnosis of tumors, tumor staging, typing, early detection of tumor metastasis and prognosis. However, we also see that Real-Time PCR detects false positives and false negatives, but believes that with the advancement of technology, Real-Time PCR technology is used to jointly detect multiple genes for comprehensive index analysis, or Other technologies, such as reverse transcription PCR, immunohistochemistry, HE staining, etc., make Real-Time PCR technology a broader application prospect in clinical and more fields.
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